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In this review we analyze the multi-faceted role that B cells play in the pathogenesis of rheumatoid arthritis and discuss how this information can be used to guide more specific targeti

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There is significant evidence arising from experimental models that

autoantibodies play a key role in the pathogenesis of inflammatory

arthritis In addition to autoantibody production, B cells efficiently

present antigen to T cells, produce soluble factors, including

cyto-kines and chemocyto-kines, and form B cell aggregates in the target

organ of rheumatoid arthritis In this review we analyze the

multi-faceted role that B cells play in the pathogenesis of rheumatoid

arthritis and discuss how this information can be used to guide

more specific targeting of B cells for the therapy of this disease

Introduction

The advent of biological therapy has provided a powerful tool

to improve our understanding of the pathogenesis of disease

As well as benefiting patients, the success of targeted

biological therapies demonstrates the importance of particular

molecules or cells in disease pathogenesis The potent

efficacy of rituximab, a B-cell depleting agent, in the treatment

of patients with rheumatoid arthritis (RA) has revitalized

interest in the central role played by B cells in disease

pathogenesis [1] (Fig 1)

Accumulation of B cells in the synovium is

driven by a variety of signals

RA is one of only a few diseases in which ectopic germinal

centre-like structures can be observed at the site of

inflammation [2] These structures, which range from loose

aggregates of T and B cells to distinct follicle-like structures,

are often observed in close contact with the inflamed synovial

membrane of RA patients A variety of cells, including

fibro-blast-like synoviocytes and dendritic cells, that are present in

the synovium of patients with RA produce factors that affect

B-cell survival, organization and trafficking, such as B

cell-activating factor of the TNF family (BAFF), CXC chemokine

ligand (CXCL)13, CXCL12 and lymphotoxin beta (Table 1)

[2-4] Based on their immunological function and location, each of these factors could contribute to the recruitment and maintenance of B cells in arthritic joints, thus representing potential therapeutic targets For example, blockade of surface lymphotoxin using a decoy lymphotoxin receptor-immunoglobulin and BAFF, as discussed below, are currently

in clinical trials Interestingly, the efficacy observed in RA patients treated with etanercept, which binds to

lymphotoxin-α as well as tumour necrosis factor (TNF)-lymphotoxin-α, may partly be related to blockade of the former cytokine [5] Simultaneous blockade of more than one factor that drives B-cell accumulation may be a more efficient therapeutic approach than targeting a single cytokine or chemokine

The role played by B cells in the maintenance of ectopic germinal centre-like structures, as well as in the immune response in RA synovium, has been addressed by using a humanized experimental model in which synovial tissue derived from patients was implanted into severe combined immunodeficient (SCID) mice [6] B cells were then depleted

by administration of anti-CD20 (rituximab), and T-cell responses were measured The removal of B cells led to disruption of the lymphoid-like structures and to a reduction

in T-helper (Th)1 interferon-γ producing cells, which are known to be involved in the induction and maintenance of the proinflammatory cytokine cascade

Role played by B cells as antigen-presenting cells in rheumatoid arthritis

B cells actively participate in an autoimmune process through interaction with T cells by a variety of mechanisms, including antigen presentation and cytokine production B cells process antigens, which are presented to T cells via major histocompatibility complex class II Inherited susceptibility to

RA has been associated with DRB1 genes that encode the

Review

Cells of the synovium in rheumatoid arthritis

B cells

Claudia Mauri and Michael R Ehrenstein

Centre for Rheumatology Research, Department of Medicine, University College London, Cleveland Street, London, W1T 4JF, UK

Corresponding authors: Claudia Mauri, c.mauri@ucl.ac.uk, or Michael R Ehrenstein, m.ehrenstein@ucl.ac.uk

Published: 5 March 2007 Arthritis Research & Therapy 2007, 9:205 (doi:10.1186/ar2125)

This article is online at http://arthritis-research.com/content/9/2/205

© 2007 BioMed Central Ltd

RA = rheumatoid arthritis; BAFF = B cell-activating factor of the TNF family; NP = hapten 4-hydroxy-3-nitro-phenyl acetyl; CCP = cyclic citrullinated peptide; CFA = complete Freund’s adjuvant; CIA = collagen-induced arthritis; CXCL = CXC chemokine ligand; FcγR = Fcγ receptor; IL = inter-leukin; IVIG = intravenous IgG; PG = prostaglandin; RF = rheumatoid factor; SCID = severe combined immunodeficient; Th = T-helper (cell); TLR = Toll-like receptor ligand; TNF = tumour necrosis factor;

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HLA-DR4 and HLA-DR1 molecules [7] These findings

suggest a pathogenic role for antigen presentation in RA

Although dendritic cells are believed to be important in

priming nạve T cells, B cells represent the predominant

population of antigen-presenting cells in later phases of the

immune response [8] Rheumatoid factor (RF)-producing B

cells are particularly effective in presenting immune

complexes to T cells, irrespective of the antigen contained in

the antigen-antibody complex [9] Thus, T cells of other

specificities could easily be activated if immune complexes in

RA contain other antigens

T-cell priming by B cells has been shown to be important in the

pathogenesis of a murine model of arthritis Specifically, the

involvement of B cells in priming T cells was dissected within

the context of proteoglycan (PG)-induced arthritis by using a

mouse deficient in a secretory antibody (mIgM) [10] These

mice express a membrane-bound heavy chain transgene,

which pairs with an endogenous light chain specific for hapten 4-hydroxy-3-nitro-phenyl acetyl (NP) T cells isolated from PG-immunized mIgM mice failed to induce arthritis in SCID mice, even if they were co-transferred with wild-type B cells, suggesting that T cells do not become properly primed in this experimental setting However, targeting PG to B cells using

NP coupled with PG led to differentiation of arthritogenic T cells that are able to transfer disease Other antigen-presenting cells could not substitute for B cells in this T-cell priming, supporting a central role for B cells in driving autoreactive T cells Autoantibody production was also essential for development of severe disease, indicating that B cells play two complementary roles in the pathogenesis of arthritis

Immune complexes can activate B cells via Toll-like receptor ligands

It was recently shown that chromatin-containing immune complexes can activate B cells through Toll-like receptor

Figure 1

B cell participation in RA Illustrated is the potential role of B cells in the regulation of immune responses in RA Mature B cells, upon antigen encounter and TLR stimulation, expand and differentiate into short-lived plasma cells or can enter into a GC reaction, which is necessary for the generation of both memory B cells, and long-lived plasma cells that can produce autoantibodies Autoantibodies form immune complexes that further activate the immune system via Fc and complement receptors expressed on target cells Antigen-activated mature B cells provide help to T cells and induce differentiation of effector T cells that produce proinflammatory cytokines (known to be directly/indirectly involved in cartilage and bone destruction) Mature B cells, via mechanisms yet to be elucidated, can also differentiate into IL-10 producing B cells that can dampen the autoreactive T-cell response GC, germinal centre; IFN, interferon; IL, interleukin; RA, rheumatoid arthritis; TLR, Toll-like receptor ligand; TNF, tumour necrosis factor

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ligand (TLR)9 These immune complexes activate B cells to

produce RF by synergistic engagement of B-cell receptor

and TLR9 [11] TLRs were originally described as a family of

pattern-recognition receptors that can differentiate between

microbial molecular patterns and host components [12] Their

engagement results in rapid activation of the innate and

adaptive immune systems to effect clearance of pathogens

There is mounting evidence suggesting involvement of TLR

signalling in the pathogenesis of experimental arthritis Mice

deficient for MyD88, the essential adaptor molecule involved

in signalling by TLR family members, failed to develop

streptococcal cell wall induced arthritis, and TLR2-deficient

mice exhibited reduced disease [13] Furthermore, direct

injection of CpG DNA or double-stranded RNA into joints of

susceptible mice results in the development of transient

arthritis [14] Heat shock protein, fibrinogen and hyaluronan,

which are known to bind to TLR4, have all been detected in

the inflamed joint [15] In the KB×N model of murine antibody

transferred arthritis, TLR4-deficient mice exhibit reduced

disease [16] Although there is enough evidence from

experimental arthritis implicating TLRs in the development of

arthritis, whether TLR activation is involved in human RA

remains to be formally demonstrated

Autoantibodies as effector molecules in

rheumatoid arthritis

The pathological involvement of antibodies in inflammatory

arthritis was first proved using DBA/1 mice immunized with

collagen type II in complete Freund’s adjuvant (CFA) These

mice develop a severe arthritis that shares some pathological

features with human RA It has been demonstrated that SCID

mice (which lack T and B cells), when treated with serum

isolated from arthritic DBA/1 mice, develop an inflammatory

arthritis [17,18] However, the disease was transient and less severe than collagen-induced arthritis (CIA) A more severe arthritis can be induced in recipient mice if serum is cotransferred with T cells presensitized with heat denatured collagen [19]

The pathological relevance of B cells in arthritis was further demonstrated by Holdhmal and colleagues [20], using µMT mice (which lack B cells) immunized with type II collagen in CFA Lack of B cells completely prevented induction of arthritis and resulted in an impaired T-cell response to type II collagen (Mauri C, unpublished data); this suggests that although autoantibodies can initiate disease, other compo-nents of the immune system are needed to fuel the pathogenic response

Compared with evidence from experimental models of arthritis, proof that antibodies are also pathogenic in human

RA is more difficult to obtain The presence of RF in serum, which binds to the constant region of IgG, was first identified

in 1957 [21] and has long been recognized as a marker in the majority of patients with RA The severity of RA has been correlated with RF levels, and patients who are seropositive for RF have more aggressive disease and worse prognosis [22,23] Important studies have indicated that the presence

of RF can be detected many years before arthritis begins [24,25]

The possible pathogenic role of RF-positive B cells has been revisited with the increased use of rituximab in RA therapy RF can cause tissue damage through formation of immune complexes, by activation of complement, thereby recruiting cells into the synovium Although preliminary data suggested that patients who are RF negative appear less likely to respond to B-cell depletion therapy [26], a larger trial [27] did not identify substantial differences in the response between RF-positive and RF-negative patients with RA However, RF titres fell by 55% in those patients who were RF positive, which corroborates previous findings [28] Moreover, routine assays for RF are not particularly sensitive and do not exclude their presence Thus, patients identified as being RF-negative may still have low titres of RF It is also possible that the many other autoantibody specificities present in patients with RA may distinguish those patients who respond to rituximab These other autoantibody specificities include those directed toward the nuclear antigen RA-33 and heavy chain binding protein, both of which are found in early RA and pre-disease sera [29] Antibody and T-cell reponses to heavy chain binding protein have been identified both in patients with RA and in animal models, suggesting that this may be an important autoantigen Although antibodies to type II collagen have been shown to induce disease in animal models, it is unlikely that anti-collagen antibodies are relavant to human disease Recently, antibodies to citrulline-modified peptides (anti-cyclic citrullinated peptide [CCP] antibodies) have attracted

Table 1

Potential targets driving B cell accumulation in the synovium

CXCL12 (SDF-1) B cell and plasma cell chemoattractant

CXCL13 B cell chemoattractant

Lymphotoxin-β Organization of B cells within lymphoid

architecture Induction of CXCL13

IL-5, IL-6, TNF-α Promote plasma cell accumulation and

survival

BAFF Immature and mature B cell survival and

proliferation

APRIL Mature B cell and plasma cell survival

TNF-α and IFN-γ Induces B cell release from bone marrow

Increased production of BAFF and APRIL

APRIL, a proliferation-inducing ligand; BAFF, B cell-activating factor of

the TNF family; CXCL, CXC chemokine ligand; IFN, interferon;

IL, interleukin; TNF, tumour necrosis factor

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considerable attention, and their measurement has now

entered into routine clinical use From a clinical perspective,

anti-CCP antibodies represent a useful test for predicting

which patients with early arthritis will go on to develop RA

The presence of both anti-CCP antibodies and RF predicts

the development of RA in patients with early arthritis with

high sensitivity and specificity [25] The importance of

anti-CCP antibodies is further emphasized by their link with

HLA-DRB1 shared epitope alleles, the most important genetic risk

factor for RA Recent work has suggested that this HLA

genetic risk factor is linked to the development of anti-CCP

antibodies rather than to the disease itself [30] Thus, the

presence of these autoantibodies in RA, often preceding

disease by many years, may indicate a breakdown in central

and/or peripheral tolerance

The pathological role played by anti-CCP antibodies was

recently confirmed in the CIA arthritis model As in human RA,

anti-CCP antibodies can be detected before the onset of

disease and are present in inflamed synovium of mice in the

acute phase of CIA Although the amount of anti-CCP

antibodies measured in serum of mice with acute

inflammation is similar to levels of antibodies to collagen type

II, transfer of anti-CCP antibodies alone failed to induce

disease in recipient mice However, transfer of anti-CCP

antibodies to SCID mice significantly reduced the amount of

anti-collagen type II antibodies necessary to induce disease,

demonstrating a contributing role in the development of

arthritis [31] The relevant targets of these antibodies in joints

or in peripheral tissue remain unknown

Are both Fc receptors and complement

component C5a required for autoantibodies

to drive the effector phase in arthritis?

Antibodies can act directly on target organs and induce

disease through Fc-mediated activation of the complement

system or through the formation of immune complexes In

addition, antibodies can directly activate Fcγ receptors

(FcγRs) expressed on both myeloid and lymphoid cells A

clear insight into how antibodies work in arthritis has been

gleaned from the K/B×N model of RA In these mice

glucose-6-phosphate isomerase (GPI) is the target autoantigen, and

T-cell reactivity to this ubiquitous antigen results in

recruitment of anti-GPI B cells and subsequent immune

complex-mediated arthritis [32,33] Arthritis can be induced

in non-autoimmune recipients, or in RAG2-/-mice (which lack

both T and B cells), by transfer of sera or purified antibodies

Similar to the SCID model mentioned above, inflammation

begins to subside between 15 and 30 days after antibody

transfer Histologically, analysis of joints 30 days after transfer

revealed less inflammation than in the K/B×N model itself,

and little cartilage damage, supporting the notion that other

abnormalities in cellular types and soluble factors are needed

for full expression of disease [32] FcγRs are intimately

involved in the pathogenesis of this arthritis In particular,

much milder arthritis was observed in mice lacking the FcγRIII

receptor [34], whereas FcγRII-deficient mice exhibited acce-lerated disease

The involvement of FcγRIIB in mediating antibody damage has also been investigated in the CIA model of arthritis FcγRIIB is an inhibitory receptor that suppresses B cells, mast cells and macrophages, and transmits its inhibitory signal via its immunoreceptor tyrosine-based inhibitory motif Deletion of FcγRIIB renders DBA/1 mice more susceptible to disease [35] Recently, the pathogenicity of human RA-associated antibodies was also tested in a passive transfer model using FcγRIIB deficient mice Transfer of serum from active RA patients, or an immunoglobulin-rich fraction, to 8- to 12-week-old B6.FcγRIIB-/- mice induced a mild transient arthritis [36], indicating that serum from patients with RA can induce an inflammatory arthritis The administration of a large amount of intravenous IgG (IVIG) is a common treatment for a number of autoimmune conditions and is thought to modulate

Fc receptor function [37] IVIG has been shown to have a protective effect in the K/B×N mouse model of arthritis discussed above through induction of FcγRIIB [38] This property of IVIG has been linked to sialylation of the Fc portion of IgG The proportion of sialylated IgG molecules in commercial IVIG may account for the very mixed results obtained when patients with RA were treated with IVIG [39] Involvement of the complement system in the development of mouse models of arthritis caused by autoantibodies has been demonstrated using C5-deficient mice Both the K/B×N and collagen-induced model of arthritis are dependent on C5a for disease expression, and antibodies to C5 ameliorated disease in the K/B×N model [40,41] This has led to clinical trials of C5a receptor based peptides in RA, with mixed results Other components of the complement system such

as C4 do not participate in disease pathogenesis [40] Therefore, the effector function of arthritogenic antibodies rely both on Fc receptors and C5a

Immunoregulation by B cells

Although the pathogenic role played by mature B cells in RA has been extensively studied, new data have demonstrated that a distinct subset of B cells, namely those that produce IL-10, are involved in the downregulation of the immune system It was originally demonstrated that B-cell-deficient mice developed an exacerbated experimental autoimmune encephalomyelitis as compared with wild-type animals, suggesting a protective role for B cells in the development of autoimmune disease [42] B cells that produce cytokines, and in particular IL-10, have been reported to play an immunoregulatory role in autoimmunity, chronic inflammatory bowel disorders, asthma and infectious diseases [43,44] In the context of arthritis, we previously showed that stimulation

of splenic B cells isolated during the acute phase of disease, with an agonistic anti-CD40 antibody, induces differentiation

of IL-10 producing B cells Transfer of anti-CD40 stimulated

B cells to DBA/1 mice immunized with collagen type II in CFA

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prevented or ameliorated arthritis [45] The mechanisms by

which this subset of B cell regulates the immune response

against autoantigens are not fully understood However, we

showed that mice treated with anti-CD40 challenged B cells

exhibit an impaired Th1 response [45] Therefore one

plausible explanation is that production of IL-10 might restore

the dysregulated Th1/Th2 balance, or it could directly

modulate effector cells, including macrophages and dendritic

cells, thus downmodulating inflammatory responses IL-10

producing B cells could also act as secondary

antigen-presenting cells, leading to an abortive response and

induction of anergic CD4+ T cells, or they could recruit

regulatory T cells or induce their differentiation If an

equivalent population exists in humans, then removal of these

B cells by rituximab might be detrimental

Targeting bad B cells

An understanding of which B cells are relevant to the

pathogenesis of disease is important in designing therapeutic

strategies to target B cells The vast majority of B cells found

in peripheral blood are removed by rituximab, but the extent

and nature of B cell removal in other tissues in RA patients

remain to be established For example, experiments

conducted in monkeys revealed that B cells residing in

tissues are less effectively removed, and that memory B cells

are more resistant to depletion than nạve ones [46] Similar

observations have been made in murine studies using

anti-CD20, where marginal zone B cells, B1 cells and germinal

centre B cells are more resistant to depletion [47-49] As

mentioned above, it is likely that plasma cells that producing

RF are likely to be important in disease pathogenesis, but

their depletion by rituximab is hampered by lack of CD20

expression However, those plasma cells that have a short life

span rely on CD20-expressing B cell precursors for

continued renewal Examination of peripheral blood indicates

that CD19+CD20-plasmablasts decrease following rituximab

therapy [50] The observation that RF titres decrease

following rituximab treatment suggests that short-lived, rather

than long-lived, plasma cells are at least partly responsible for

their formation A number of factors are known to be

important in plasma cell survival, including cytokines such as

TNF-α and the cell adhesion molecule CD44 [51] Perhaps

use of anti-TNF-α together with rituximab may have

syner-gistic benefit through their combined targeting of B cells and

plasma cells, although infection-related side effects may

prohibit use of this combination

Antagonists to BAFF also lead to incomplete removal of

peripheral and lymphoid B cells in monkeys, with marginal

zone-like B cells being particularly susceptible to depletion

[52] Trials have begun to evaluate the anti-BAFF agent

belimumab (LymphoStat-B; Human Genome Sciences,

Rockville, MD, USA) in RA and have demonstrated limited

efficacy, perhaps because of incomplete blockade or

because other related B-cell survival factors such as a

proliferation-inducing ligand (APRIL) would not be affected

Conclusion

Renewed interest in B cells in RA has been initiated by a global B-cell-depleting agent, but it is likely that only a small proportion of B cells contribute to disease pathogenesis whereas others may actually be protective It is hoped that research in patients with RA using these new agents will reveal correlations between pathogenic B cell subsets and improvement in clinical disease activity, thereby enhancing our understanding of the role played by B cells in human disease

Competing interests

The authors declare that they have no competing interests

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